New insights on the phylogeography of Hepatozoon canis in Brazil.

This study aimed to molecularly characterize the Hepatozoon spp. infecting domestic and wild dogs in Brazil. A total of 22 whole blood samples tested positive for Hepatozoon spp., and five samples were sequenced for the 18S rDNA gene from H. canis after PCR amplification with four primer sets. Phylogenetic analysis using Bayesian inference showed that the three H. canis isolates from domestic dogs were not monophyletic; however, they were more closely related to each other than to other H. canis sequences. The isolate from the hoary fox (Lycalopex vetulus) was phylogenetically more distant. Two haplotype networks were constructed, identifying 10 haplotypes of H. canis in Brazil, with H10 constituting the largest group. It contains nine isolates, including three from domestic dogs. The H5 haplotype grouped the sequence of L. vetulus with two additional sequences from hosts Tapirus terrestris and L. vetulus, representing the sole haplotype with wild hosts. Bayesian analysis suggested the possible existence of two genetic groups of H. canis in Brazil, indicating gene flow of this agent within the country. These findings contribute valuable insights for a more comprehensive understanding of the molecular diversity of Hepatozoon spp. in Brazil and may help in the development of effective control measures.

Main mites associated with dermatopathies present in dogs and other members of the Canidae family.

Dermatological diseases of parasitic origin are one of the most frequent in the clinical practice of dogs and cats. Mites such as Sarcoptes scabiei, Otodectes cynotis, Demodex canis, and Cheyletiella spp., commonly affect domestic dogs. However, the impact generated by these mites on populations of wildlife animals and the mechanisms involved in their epidemiological dynamics are still not clear. In recent decades, the migration of populations and their interaction with domestic environments and vice versa have generated a worrying threat due to the transmission of some of these ectoparasites. Some reports have suggested that sarcoptic mange represents an emerging threat to wildlife. Given the outbreaks of greater magnitude and geographical extension. The objective of this review is to contribute to the state of the art of the main mites that cause dermatopathies in members of the Canis lupus familiaris family and other members of the Canidae family. For this, a systematic search was carried out in the Embase and PubMed databases. Infections caused by mites, mainly scabies, continue to be diseases with a worldwide distribution, affecting mammals and humans. Although they are long-standing diseases, the effects that are generated in wild canids are still unknown. A comprehensive evaluation is required to generate guidelines in favor of the conservation of some species of foxes and wolves present in different regions of the world.

The Power of Discourse: Associations between Trainers' Speech and the Responses of Socialized Wolves and Dogs to Training.

In a previous study, we found that Positive Reinforcement Training reduced cortisol of wolves and dogs; however, this effect varied across trainer-animal dyads. Here we investigate whether and how the trainers' use of speech may contribute to this effect. Dogs' great interest in high-pitched, intense speech (also known as Dog Directed Speech) has already been reported, but whether and how wolves respond similarly/differently to voice characteristics has never been studied before. We analyzed 270 training sessions, conducted by five trainers, with nine mixed-breed dogs and nine wolves, all human-socialized. Through Generalized Linear Mixed Models, we analyzed the effects of (a) three speech categories (nice, neutral, reprehensive) and laugh; and (b) acoustic characteristics of trainers' voices on animals' responses (correct responses, latency, orientation, time at less than 1 m, non-training behaviors, tail position/movements, cortisol variation). In both subspecies, tail wagging occurred more often in sessions with longer durations of nice speech, and less often in sessions with reprehensive speech. For dogs, the duration of reprehensive speech within a session was also negatively related to correct responses. For wolves, retreat time was associated with more reprehensive speech, whereas duration of nice speech was positively associated with time spent within one meter from the trainer. In addition, most dog behavioral responses were associated with higher average intonations within sessions, while wolf responses were correlated with lower intonations within sessions. We did not find any effects of the variables considered on cortisol variation. Our study highlights the relevance of voice tone and speech in a training context on animals' performances and emotional reactions.

Outbreak of canine distemper and coinfections in a maned wolf (Chrysocyon brachyurus) and in three giant anteaters (Myrmecophaga tridactyla).

Canine distemper outbreak and coinfections in three giant anteaters and in a maned wolf has been described. Three giant anteaters developed respiratory and digestive clinical signs after the introduction of a maned wolf to a Wildlife Rehabilitation Center. The maned wolf and two anteaters died, and one anteater was euthanized. Post mortem and histopathologic exams revealed lesions associated with numerous intraepithelial inclusion bodies, mainly in the respiratory and digestive systems. Infection by distemper virus was confirmed in all animals by RT-PCR and gene sequencing, which revealed the Europe 1/ South America 1 strain, closely related to the strain from Canis familiaris. In addition to distemper, the animals had other comorbidities, such as toxoplasmosis and salmonellosis in the maned wolf and cutaneous candidiasis in an anteater. Considering the chronology of clinical manifestation in both species and the viral characterization, it is possible that the maned wolf was the source of infection to the anteaters. This study demonstrates the importance of implementing biosecurity measures in enclosures that house animals of different species, highlighting the importance of quarantine before introduction of new animals into the same environment.

Cinomose em canídeos silvestres no Brasil / Distemper in wild canids in Brazil

A cinomose é uma doença viral multissistêmica causada por um Morbillivirus. No Brasil, existem seis espécies de canídeos silvestres vulneráveis a essa enfermidade. Dessa forma, este trabalho tem como objetivo apresentar uma revisão sobre a situação da cinomose em canídeos silvestres no Brasil e os impactos causados na fauna brasileira. A transmissão do vírus ocorre através do contato com amostras contaminadas, a exemplo dos aerossóis, excretas e secreções de indivíduos infectados. Os sinais clínicos mais prevalentes são de ordem neurológica, entretanto, também podem ser identificadas manifestações respiratórias, cutâneas e gastrointestinais. Portanto, o diagnóstico consiste na avaliação da sintomatologia apresentada em conjunto com testes específicos, como isolamento viral, ELISA, imunofluorescência e RT-PCR. Atualmente, não existe tratamento específico. Desta forma, são realizados apenas cuidados paliativos. Os grandes carnívoros são os mamíferos mais ameaçados do mundo, sobretudo, em consequência dos impactos causados pela redução do habitat natural, associada à expansão territorial humana. As epidemias também justificam as altas taxas de mortalidades desses animais, o que pode estar relacionado com comportamentos sociais e de farejamento, assim como o crescente contato entre animais silvestres e domésticos devido à urbanização e à proximidade genética que os canídeos silvestres têm com os cães domésticos, tornando-os suscetíveis às infecções. Visto que a cinomose é uma patologia emergente em populações de carnívoros silvestres e que a presença de cães domésticos não vacinados em áreas de conservação representa um grande risco de contaminação, conclui-se que a não vacinação está diretamente associada à perpetuação do vírus no meio selvagem.

Distemper is a multisystem viral disease caused by a Morbillivirus. In Brazil, there are six species of wild canids, vulnerable to this disease. Therefore, the present work aims to develop a review on the situation of distemper in wild canids in Brazil and the impacts caused on the Brazilian fauna. The virus transmission occurs through contact with contaminated samples, such as aerosols, excreta, and secretions from infected individuals. The most prevalent clinical signs are neurological; however, respiratory, cutaneous and gastrointestinal manifestations can also be identified. Thus, the diagnosis consists of evaluating the symptoms presented together with specific tests, such as viral isolation, ELISA, immunofluorescence, and RT-PCR. Currently, there is no specific treatment. Therefore, only palliative care is performed. Large carnivores are the most threatened mammals in the world, mainly as a result of the impacts caused by the reduction of natural habitat, associated with human territorial expansion. Epidemics also justify the high mortality rates of these animals, which may be related to social and sniffing behaviors, as well as the increasing contact between wild and domestic animals due to urbanization and the genetic proximity that wild canids have with domestic dogs, making them susceptible to infections. Since distemper is an emerging pathology in populations of wild carnivores and the presence of unvaccinated domestic dogs in conservation areas represents a great risk of contamination, it is concluded that non-vaccination is directly associated with the perpetuation of the virus in the wild.

Chagas Immunochromatographic Rapid Test in the Serological Diagnosis of Trypanosoma cruzi Infection in Wild and Domestic Canids.

Canis lupus familiaris (domestic dog) represents a reliable sentinel for the occurrence of a well-established transmission cycle of Trypanosoma cruzi among wild mammals in the surroundings and, consequently, where the risk of human infection exists. Serological diagnosis is the chosen method to identify T. cruzi infection in dogs that, in Brazil, rarely present positive parasitological tests. The use of recombinant chimeric parasitic antigens results in a sensitive and specific serological diagnostic test in contrast to the use of crude T. cruzi antigens. Our objective was to evaluate the Chagas/Bio-Manguinhos Lateral Flow Immunochromatographic Rapid Test (Chagas-LFRT) for the diagnosis of T. cruzi infection in domestic dogs and the potential of application of this diagnostic platform to wild canid species. Two recombinant proteins (IBMP-8.1 and IBMP-8.4) that displayed the best performance in the enzyme immunoassay (ELISA) in previous studies were tested in a platform with two diagnostic bands. A panel of 281 dog serum samples was evaluated: 133 positive for T. cruzi by serological diagnosis, including 20 samples with positive blood cultures belonging to different discrete typing units (DTUs); 129 negative samples; and 19 samples from dogs infected by other trypanosomatids: Leishmania infantum, Trypanosoma rangeli, Trypanosoma caninum and Crithidia mellificae, in addition to samples infected by Anaplasma platys, Dirofilaria immitis and Erlichia sp. that were employed to evaluate eventual cross-reactions. We also evaluated the Chagas-LFRT to detect T. cruzi infection in 9 serum samples from six wild canid species. We observed that the intensity pattern of the bands was directly proportional to the serological titer observed in IFAT. The sensitivity was 94%, the specificity was 91% according to the ROC curve, and the defined cutoff was an optical density of 4.8. The agreement obtained was considered substantial by the kappa analysis (84%). From T. cruzi positive hemoculture samples, 88.9% were positive by Chagas-LFRT. The test was efficient in recognizing infections by five of the six T. cruzi DTUs. Cross-reactions were not observed in infections by L. infantum, T. rangeli, T. caninum and D. immitis; however, they were observed in sera of dogs infected by Crithidia mellificae, Anaplasma sp. and Erlichia sp. A strong reaction was observed when serum samples from wild canids were submitted to the Protein A affinity test, confirming its applicability for these species. This test will allow rapid preventive actions in areas with high risk to the emergence of Chagas disease in a safer, reliable, low-cost and immediate manner, without the need for more complex laboratory tests.

Alimentação natural em cães da raça Dobermann: estudo bioquímico sérico / Natural food in Dobermann breed dogs: serum biochemical study

O objetivo desse estudo foi determinar o perfil bioquímico sérico de cães da raça Dobermann que consomem alimentação natural. Foram realizados exames de albumina, proteínas totais, colesterol total, triglicerídeos, aspartato aminotrasnferase, alanina transaminase, fosfatase alcalina, glutamiltransferase, ureia, ácido úrico, lipase, glicose, creatinina, desidrogenase láctica e creatina quinase. As principais alterações incluem a hiperalbunemia, aumento de triglicerídeos, desidrogenase láctica, ureia e lipase. A alimentação natural influencia em parâmetros bioquímicos, mostrando a importância de se determinar valores de referência para tal prática. Além disso, este trabalho é um dos primeiros que mostra a efetividade da alimentação natural em cães da raça Dobermann.

The aim of this study was to determine the serum biochemical profile of Dobermann dogs that consume natural food. Albumin, total proteins, total cholesterol, triglycerides, aspartate aminotransferase, alanine transaminase, alkaline phosphatase, glutamyltransferase, urea, uric acid, lipase, glucose, creatinine, lactic dehydrogenase and creatine kinase tests were performed. The main alterations include hyperalbunemia, increase in triglycerides, lactic dehydrogenase, urea and lipase. Natural food influences biochemical parameters, showing the importance of determining reference values for this practice. In addition, this work is one of the first to show the effectiveness of natural feeding in Dobermann dogs.

Colonic papillary adenocarcinoma in a crab-eating fox (Cerdocyon thous): Anatomopathological and immunohistochemical features

A captive 17-year-old male crab-eating fox (Cerdocyon thous) presenting clinical signs of hyporexia and constipation for two weeks was found dead. On autopsy, a large and firm mass expanding the large intestine wall and totally obstructing the colon lumen was identified. Histopathology revealed a non-encapsulated poorly demarcated adenocarcinoma composed of cuboidal to pseudostratified columnar epithelium forming papillary projections of variable sizes. The cytoplasm of the neoplastic cells was immunopositive for CK7 and PAN-CK, but negative for VIM and CK20. Immunohistochemistry revealed an immunophenotype different from the enteric adenocarcinomas reported in humans and domestic dogs. This is the first case report of colonic papillary adenocarcinoma in a crab-eating fox.(AU)

The importance of the dog (Canis lupus familiaris) in cocoa farms as carriers of helminths potentially transmissible to humans and wildlife in the Southern Bahia, Brazil / A importância do cão (Canis lupus familiaris) nas fazendas de cacau como portador de helmintos potencialmente transmissíveis ao homem e à vida selvagem no sul da Bahia, Brasil

Domestic dogs (Canis lupus familiaris) can be considered an invasive species as they have been introduced by humans in different parts of the world and represent a risk to wildlife conservation. Large extensions of agroforestry systems, where cocoa is grown under the shade of native trees, contribute to wildlife conservation in southern Bahia, Brazil. However, this system can increase contact between species of the native fauna and domestic dogs, which are frequently taken to the fields by rural workers. The aims of this study were to investigate the presence of domestic dogs inside cocoa agroforestry systems and the occurrence of helminths in dogs from cocoa farms near two protected areas: the Una Biological Reserve (REBIO-Una) and the Serra das Lontras National Park in Una, Bahia. We also investigated general characteristics of the dogs such as age, sex, breed, feeding frequency, vaccination, deworming and others, and evaluated possible risks of wildlife and human contamination by the domestic dog parasites. Camera traps were set up on eight cocoa agroforestry systems and three adjacent forest fragments. For parasitological investigation, fecal samples were collected from domestic dogs that lived on the eight cocoa farms. A total of 539 photos of 12 mammals were registered, of which 15% were of domestic dogs. The parasitological research was carried out with fecal samples from 32 of the 39 dogs that lived in the cocoa farms. We found one genus of cestode, Dipylidium sp. (7.7%), and five genera of nematodes, Ancylostoma sp. (80.7%), Strongyloides sp. (38.4%), Toxocara sp. (30.7%), Spirocerca sp. (15.4%) and Trichuris sp. (11.5%). Regarding animal care, all dogs were in poor body condition, 49% were fed twice a day, 90% defecated in the forest or cocoa agroforestry systems, only 33% were periodically dewormed, 64% were vaccinated against rabies in less than 12 months, and only one dog was vaccinated against other viruses. The results showed that domestic dogs share the same environment as wild animals yet lack some basic health care, increasing the possibility of parasitic transmission between domestic dogs, wildlife and humans from environmental contamination. We highlight the need for responsible pet ownership and the monitoring the health of wild animals and the human population living in this region.

Cães domésticos (Canis lupus familiaris) podem ser considerados espécies invasoras introduzidas pelo ser humano em diversos lugares do mundo, representando risco para a conservação da fauna. Grandes extensões de sistemas agroflorestais, onde o cacau é cultivado sob a sombra de árvores nativas, contribuem para a conservação da vida selvagem no sul da Bahia, Brasil. Entretanto, esse sistema pode aumentar o contato entre espécies da fauna nativa e cães domésticos, frequentemente levados para o campo por trabalhadores rurais. Os objetivos deste estudo foram investigar a presença de cães domésticos em sistemas agroflorestais de cacau e a ocorrência de helmintos em cães de fazendas de cacau nas proximidades de duas áreas protegidas: a Reserva Biológica de Una (REBIO-Una) e o Parque Nacional da Serra das Lontras, em Una, Bahia. Também foram investigadas as características gerais dos cães como idade, sexo, raça, frequência de alimentação, vacinação, vermifugação e outros, e avaliados os possíveis riscos de contaminação da vida selvagem e seres humanos por parasitas de cães domésticos. Armadilhas fotográficas foram instaladas em oito sistemas agroflorestais de cacau e três fragmentos florestais adjacentes. Para investigação parasitológica, foram coletadas amostras fecais de cães domésticos que viviam nas oito fazendas de cacau. Foram registradas 539 fotos de 12 mamíferos, onde 15% das fotos pertenciam à cães domésticos. A pesquisa parasitológica foi realizada com amostras fecais de 32 dos 39 cães que viviam nas fazendas de cacau. Encontramos um gênero de cestóide: Dipylidium sp. (7,7%), e cinco gêneros de nematóides: Ancylostoma sp. (80,7%), Strongyloides sp. (38,4%), Toxocara sp. (30,7%), Spirocerca sp. (15,4%) e Trichuris sp. (11,5%). Em relação aos cuidados com os animais, todos estavam com score corporal baixo, 49% dos cães eram alimentados duas vezes ao dia, 90% defecam na floresta ou em sistemas agroflorestais de cacau, apenas 33% foram vermifugados periodicamente, 64% vacinados contra a raiva em menos de 12 meses e apenas um cão foi vacinado contra outros vírus. Os resultados mostram que os cães domésticos compartilham o mesmo ambiente que os animais silvestres e carecem de alguns cuidados básicos de saúde, aumentando a possibilidade de trânsito parasitário entre cães domésticos, animais silvestres e humanos por contaminação ambiental. Destaca-se a necessidade de atividades de posse responsável de animais de estimação e o monitoramento da saúde dos animais silvestres e da população humana que vive nesta região.

Biotécnicas reprodutivas: captura, exame andrológico, conservação do ejaculado e inseminação artificial em canídeos e felídeos selvagens em risco de extinção / Reproductive biotechniques: capture, andrological examination, ejaculate conservation and artificial insemination in wild dogs or felids in extinction risk / Biotecnicas reproductivas: captura, examen andrológico, conservación eyaculada e inseminación artificial en cánidos e felinos silvestres en riesgo de extinción

Debido a la reducción acelerada del número de animales y la consecuente disminución de la variabilidad genética, fue necesario estudiar y aplicar biotecnologías de cría en animales silvestres y salvajes, con el fin de reducir el impacto sobre ellos y disminuir el riesgo de extinción de diferentes especies de cánidos y felinos salvajes y salvajes. El presente estudio, mediante metodología cualitativa, mediante revisión bibliográfica, realizó análisis de estudios y datos existentes, con el objetivo de dilucidar y describir los métodos de captura de los animales, el examen andrológico, los medios de preservación del eyaculado y la inseminación artificial en cánidos y felinos silvestres con el fin de preservar especies en peligro de extinción.(AU)

Due to the accelerated reduction in the number of animals and the consequent decline in genetic variability, it was necessary to study and apply breeding biotechniques in wild and wild animals, in order to reduce the impact on them and decrease the risk of extinction of different species of canids and wild and wild felids. The present study, through qualitative methodology, through bibliographic review, carried out analysis of existing studies and data, with the objective of elucidating and describing the methods of capturing the animals, the andrological examination, means of preserving the ejaculate and artificial insemination canids and wild felids for the purpose of preserving endangered species.(AU)

Devido a acelerada redução no número de animais e consequente declínio na variabilidade genética, foi necessário estudar e aplicar biotécnicas de reprodução em animais silvestres e selvagens, com a finalidade de reduzir o impacto sobre estas e diminuir o risco de extinção de diferentes espécies de canídeos e felídeos selvagens e silvestres. O presente estudo, através de metodologia qualitativa, por meio de revisão bibliográfica, realizou análise de estudos e dados existentes, com objetivo de elucidar e descrever os métodos de captura dos animais, o exame andrológico, meios de conservação do ejaculado e a inseminação artificial em canídeos e felídeos selvagens com fim de preservação das espécies ameaçadas de extinção.(AU)

Influence of extraordinary floods on wildlife parasites: the case of gastrointestinal helminths and protozoa of wild canids from the Iberá Ecoregion, Argentina.

Parasites are natural components of ecosystems and play a significant role in the dynamics of wild animal populations. Although the environment of parasites is primarily defined by the host, most life cycles involve stages that must endure external conditions. Rainfall and flooding events are important factors that might influence the transport of parasitic stages, altering soil moisture levels, and resulting in a favorable environment for parasite survival and development. We assessed whether an extraordinary flood event modified the occurrence of gastrointestinal parasites (nematodes and protozoa) in wild canids in two protected areas in northern Argentina. From 2016 to 2018, we collected fecal samples of two fox species, Lycalopex gymnocercus and Cerdocyon thous, and examined the presence of nematodes and protozoa. We assessed changes in the occurrence of these parasites after a flood event, while adjusting for potential confounders (i.e., monthly average temperature, season, host species, site). In a second stage of the analysis, we evaluated whether part of the effect was caused by changes in soil moisture, by adding normalized difference water index as an independent variable. We found that the presence of nematodes in foxes was higher after flooding than before flooding, and this association was not explained by changes in the soil moisture. On the other hand, the flood event was not relevant for protozoa. Stronger and long-lasting flood events are expected due to the effect of global warming on El Niño events, and this may increase and intensify the spread of some parasites affecting wildlife, which could also be of public health concern.

Cutaneous mast cell tumor in a captive Bush dog (Speothos venaticus): pathological and immunophenotypical aspects - case report / Mastocitoma cutâneo em um cachorro-do-mato vinagre (Speothos venaticus) de cativeiro: aspectos patológicos e imunofenotípicos - relato de caso

A senile male captive bush dog (Speothos venaticus) presented a small perianal cutaneous nodule. Histologically, there was an ulcerated round cell tumor composed of well differentiated mast cells with abundant intracytoplasmic purple Giemsa-positive granules, with a diffuse eosinophilic infiltrate. Immunohistochemistry revealed that 30% of the neoplastic cells were positive for Kit in the cytoplasm and cell membrane, and all neoplastic cells were negative for MAC and CD3. Less than 10% of the neoplastic cells were positive for Ki67. At necropsy other primary tumors were identified in this animal, including an intestinal adenoma, an adrenal cortex adenoma and a testicular interstitial cell tumor.(AU)

Um cachorro-vinagre (Speothos venaticus) apresentou um nódulo cutâneo pequeno na região perianal. Histologicamente havia neoplasia cutânea de células redondas e ulcerada, constituída por mastócitos bem diferenciados, com abundantes grânulos citoplasmáticos metacromáticos na coloração de Giemsa e infiltrado eosinofílico difuso. A imuno-histoquímica demonstrou que 30% das células neoplásicas eram positivas para a proteína Kit no citoplasma e na membrana celular. As células foram negativas para MAC e CD3. Menos de 10% das células neoplásicas foram positivas para Ki67. Durante a necropsia, foram identificados outros tumores primários, como adenoma intestinal, adenoma cortical da adrenal e tumor de células intersticiais do testículo.(AU)

Survey of Zoonotic Bacterial Pathogens in Native Foxes in Central Chile: First Record of Brucella canis Exposure.

Brucella abortus, B. canis, and pathogenic Leptospira are zoonotic pathogens that infect humans, as well as domestic and wild animals. In wild canids, they may affect their fertility and reproduction, threatening their conservation. Wild canids play a crucial role in the environment as meso- and top-predators and environmental sentinels for zoonotic pathogens. In Chile, three species of wild canids are present, and due to changes in land use and environmental dynamics, it is of utmost relevance to determine the role of these species in the epidemiology of brucellosis and leptospirosis. This study aimed to detect the exposure to B. abortus, B. canis, and pathogenic Leptospira by serologic, bacteriologic, and molecular techniques in native foxes from rehabilitation and exhibition centers in Central Chile. Forty-six blood samples were obtained from Lycalopex culpaeus and L. griseus, detecting 10.9% of seropositivity to B. canis and 7.7% to L. Javanica. No seropositivity was seen for B. abortus. Exposure was not registered by culture and qPCR in any of the sampled animals. Our findings are the first register of exposure to any Brucella species in wild canids in Chile and highlight the need to establish surveillance programs of these emerging pathogens.

Rangelia vitalii in free-living crab-eating foxes (Cerdocyon thous) in Uruguay.

Rangelia vitalii is a protozoan parasite that causes a hemorrhagic and hemolytic disease in dogs known as rangeliosis. Current reports of the disease are concentrated in the southern and southeastern regions of Brazil, as well as in Uruguay, Argentina, and Paraguay, and mainly concern domestic dogs. South American wild canids, such as the crab-eating fox (Cerdocyon thous), the pampas fox (Lycalopex gymnocercus), and the maned wolf (Chrysocyon brachyurus) may also be affected, although existing reports are restricted to Brazil. The present study aimed to detect R. vitalii parasitism in the Uruguayan wild fox population. DNA extracted from the blood and/or spleen samples of road-killed C. thous and L. gymnocercus found in northern Uruguay were subjected to polymerase chain reaction (PCR) to amplify a 551-bp fragment of the Rangelia 18S rRNA gene. A total of 62 wild canids, including 38 C. thous and 24L. gymnocercus, were analyzed. Five crab-eating fox samples (13.2%) were positive for R. vitalii, with 99.5-100% identity between the sequences. All samples from pampas fox tested negative for R. vitalii. When compared with the R. vitalii sequences available in GenBank, a similarity of 98.9-100% was revealed. Molecular analysis results suggest that R. vitalii is circulating in the crab-eating fox population in Uruguay; however, its veterinary relevance for these foxes remains unknown.

Molecular detection of coccidian Apicomplexa Parasites isolated from wild crab-eating and pampas foxes through novel TaqMan™ probes: a contribution to their molecular epidemiology.

Neospora caninum, Toxoplasma gondii and Hammondia spp. are coccidian parasites similar in morphology. Molecular techniques are necessary to detect parasite DNA isolated from stool samples in wild canids because they were reported as definitive hosts of N. caninum life cycle. The objective of this study was to develop a highly sensitive and accurate molecular method for the identification of coccidian Apicomplexa parasites in crab-eating fox (Cerdocyon thous) and pampas fox (Lycalopex gymnocercus). Tissue samples from road-killed animals (pampas fox = 46, crab-eating fox = 55) and feces (pampas fox = 84, crab-eating fox = 2) were collected, and species were diagnosed through molecular assay. PCR was used for the amplification of a fragment of the coccidian Apicomplexa nss-rRNA gene. Additionally, we developed a novel real-time PCR TaqMan™ probe approach to detect T. gondii- Hammondia spp. and N. caninum. This is the first report of N. caninum DNA in pampas fox feces (n = 1), thus it was also detected from pampas fox tissues (n = 1). Meanwhile, T. gondii was found in tissues of pampas (n = 1) and crab-eating (n = 1) foxes and H. triffittae in one crab-eating fox tissue. Despite the low percentage (2.5%) of positive samples, the molecular method developed in this study proved to be highly sensitive and accurate allowing to conduct an extensive monitoring analysis for these parasites in wildlife.

Microscopic Detection, Hematological Evaluation and Molecular Characterization of Piroplasms from Naturally Infected Dogs in Rio de Janeiro, Brazil.

PURPOSE: This study aimed to analyze the frequency of piroplasmids in the blood of dogs in Rio de Janeiro, compare the performance of microscopic techniques, assess the risk factors associated with infections and also molecularly and morphologically characterize the piroplasmids identified. METHODS: In all, 407 blood samples were collected from dogs between 2018 and 2019. These were subjected to microscopic parasitological techniques for thin and thick smears, stained with Giemsa and using a rapid staining kit. The slides were read under an optical microscope and the protozoa were characterized morphometrically. In addition, the blood samples were subjected to molecular characterization for diagnosing piroplasmid species using primers that amplified the gene 18S rRNA. RESULTS: Piroplasmids were detected in 38 (9.3%) samples. Of these, 33 samples presented nucleotide sequences compatible with Babesia vogeli. Most of the positive samples were young, male, defined breeds dogs that had been attended in clinics in São Gonçalo city. Thrombocytopenia and leukopenia were the hematological alterations more observed in positive samples, but positive samples without alterations were also detected. The sex was the only variable that showed statistical differences. Males dogs being more often infected than females (p < 0.05). The microscope slides mostly showed piriform and oval merozoites measuring greater than 2.5 µm in length, which were compatible with B. vogeli. However, smaller forms were also identified, thus demonstrating the polymorphic nature of this parasite. CONCLUSION: Babesia vogeli was detected in blood samples from dogs in the metropolitan cities of Rio de Janeiro by molecular techniques in different parasite morphotypes.

Comparison of immunohistochemical profiles of ovarian germ cells in dysgerminomas of a captive maned wolf and domestic dogs.

We characterized the immunohistochemical expression profiles of dysgerminomas from a 16-y-old maned wolf and 13 domestic dogs using the following biomarkers: Sal-like protein 4 (SALL4), octamer-binding transcription factor 3/4 (OCT3/4), placental alkaline phosphatase (PLAP), c-kit, and vimentin. The maned wolf had nonspecific and long-standing clinical signs of lethargy, anorexia, and weight loss, and was euthanized because of poor prognosis. At autopsy, the left ovary was effaced by a 12 × 8 × 6 cm mass, comprised of anaplastic cells with a mitotic count of 20 mitoses in 10 high power fields. Dysgerminomas from 7 of 13 domestic dogs had nuclear expression of SALL4. Dysgerminomas from the maned wolf and 2 domestic dogs had both nuclear and cytoplasmic expression of SALL4. Cytoplasmic expression of PLAP and OCT3/4 was present in dysgerminomas from the maned wolf and 3 (PLAP) or 4 (OCT3/4) domestic dogs. All dysgerminomas expressed vimentin. Membranous c-kit expression was rare in the dysgerminoma from the maned wolf, and variable in dysgerminomas from 4 domestic dogs. A dysgerminoma from a domestic dog had cytoplasmic expression of c-kit. SALL4 is a useful marker to confirm germ cell origin of dysgerminoma in canids.

Rangelia vitalii molecular and histological quantification in tissues comparing crab-eating foxes (Cerdocyon thous) and domestic dogs.

Rangeliosis is a condition transmitted by the tick Amblyomma aureolatum and caused by the protozoan parasite Rangelia vitalii in canids. In domestic dogs, the disease causes a severe hemolytic disease, while in wild canids the piroplasm is often detected without any clinical abnormality. This study aimed to detect and quantify the number of copies of the R. vitalii Hsp70 gene (indirect parasite burden) in several organs of domestic and South American wild canids (Cerdocyon thous and Lycalopex gymnocercus) to elucidate distinct clinical presentations of rangeliosis in these species. A total of seven domestic dogs that died due to rangeliosis and 38 wild foxes were initially included, with all dogs presenting histological and molecular features of rangeliosis, while eight C. thous were positive at the molecular analysis for R. vitalii. Fragments of 22 organs collected from domestic (n = 7) and wild foxes (n = 8) were employed for histological and molecular quantification using real-time polymerase chain reaction aiming at the Hsp70 gene. Histologically, parasitophorous vacuoles were constantly detected in the dogs, while these were detected only in two C. thous. Parasitic burden was significantly higher in the digestive, cardiorespiratory, endocrine, genitourinary, and skeletal-muscle systems of domestic dogs when compared to wild foxes. In the hematopoietic system of wild canids, some organs, such as the lymph nodes and tonsils, presented significantly lower amounts of R. vitalii, while other organs (spleen, bone marrow, and blood) had results similar to those of domestic dogs. Additionally, the central nervous system of both domestic and wild canids presented a similar quantity of R. vitalii. The etiological agent is possibly maintained through an asexual reproductive process (merogony) in both domestic and wild species. Nonetheless, a limited or short-duration schizogony phase occurs in C. thous, which would designate this species as a possible reservoir host for the agent. Dogs, in contrast, would most likely act as accidental hosts, presenting a severe and more pathogenic schizogony phase, resulting in characteristic clinical and pathological rangeliosis.

Biotécnicas reprodutivas: captura, exame andrológico, conservação do ejaculado e inseminação artificial em canídeos e felídeos selvagens em risco de extinção / Reproductive biotechniques: capture, andrological examination, ejaculate conservation and artificial insemination in wild dogs or felids in extinction risk / Biotecnicas reproductivas: captura, examen andrológico, conservación eyaculada e inseminación artificial en cánidos e felinos silvestres en riesgo de extinción

Debido a la reducción acelerada del número de animales y la consecuente disminución de la variabilidad genética, fue necesario estudiar y aplicar biotecnologías de cría en animales silvestres y salvajes, con el fin de reducir el impacto sobre ellos y disminuir el riesgo de extinción de diferentes especies de cánidos y felinos salvajes y salvajes. El presente estudio, mediante metodología cualitativa, mediante revisión bibliográfica, realizó análisis de estudios y datos existentes, con el objetivo de dilucidar y describir los métodos de captura de los animales, el examen andrológico, los medios de preservación del eyaculado y la inseminación artificial en cánidos y felinos silvestres con el fin de preservar especies en peligro de extinción.

Due to the accelerated reduction in the number of animals and the consequent decline in genetic variability, it was necessary to study and apply breeding biotechniques in wild and wild animals, in order to reduce the impact on them and decrease the risk of extinction of different species of canids and wild and wild felids. The present study, through qualitative methodology, through bibliographic review, carried out analysis of existing studies and data, with the objective of elucidating and describing the methods of capturing the animals, the andrological examination, means of preserving the ejaculate and artificial insemination canids and wild felids for the purpose of preserving endangered species.

Devido a acelerada redução no número de animais e consequente declínio na variabilidade genética, foi necessário estudar e aplicar biotécnicas de reprodução em animais silvestres e selvagens, com a finalidade de reduzir o impacto sobre estas e diminuir o risco de extinção de diferentes espécies de canídeos e felídeos selvagens e silvestres. O presente estudo, através de metodologia qualitativa, por meio de revisão bibliográfica, realizou análise de estudos e dados existentes, com objetivo de elucidar e descrever os métodos de captura dos animais, o exame andrológico, meios de conservação do ejaculado e a inseminação artificial em canídeos e felídeos selvagens com fim de preservação das espécies ameaçadas de extinção.

Spatial distribution and molecular epidemiology of hemotropic Mycoplasma spp. and Mycoplasma haemocanis infection in dogs from Rio de Janeiro, Brazil.

This cross-sectional study aims to investigate the epidemiology and spatial distribution of hemotropic Mycoplasma spp. and Mycoplasma haemocanis in dogs from Rio de Janeiro, Brazil. Blood samples were collected at random from 437 household dogs. An epidemiological questionnaire was completed concerning the host characteristics as well as the environments in which they lived. A positivity frequency of 17.84% (78/437) was found for Mycoplasma spp. and 2% (9/437) for M. haemocanis in Rio de Janeiro, Brazil, through molecular detection based on the 16S rRNA sequence. According to the present study, dogs that live in households with the presence of rodents (odds ratio [OR] = 9.93; p-value = 0.02; confidence interval [CI]: 1.34-73.66) and wild animals (OR = 1.91; p-value = 0.03; CI: 1.06-3.42) are more likely to be infected with Mycoplasma spp.. Also, dogs with tick infestation (OR = 6.47; p-value = 0.007; CI: 1.63-25.60) have more chances to become infected with M. haemocanis. The spatial analysis disclosed a positive correlation between the Mycoplasma presence and tick infestation (global Moran index = 0.82; pseudo-p-value =0.001). The epidemiological findings support the hypothesis of Rhipicephalus sanguineus s.l. as the vector of M. haemocanis in the studied region and provide insightful information to prevent the Mycoplasma spp. infection in dogs from Rio de Janeiro.